CN111500578B - Regulating ADSCs osteogenic differentiation and tissue regeneration circRNA-FTO and application thereof - Google Patents
Regulating ADSCs osteogenic differentiation and tissue regeneration circRNA-FTO and application thereof Download PDFInfo
- Publication number
- CN111500578B CN111500578B CN202010321971.1A CN202010321971A CN111500578B CN 111500578 B CN111500578 B CN 111500578B CN 202010321971 A CN202010321971 A CN 202010321971A CN 111500578 B CN111500578 B CN 111500578B
- Authority
- CN
- China
- Prior art keywords
- fto
- tissue regeneration
- circrna
- rna
- adipose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 230000017423 tissue regeneration Effects 0.000 title claims abstract description 54
- 230000009818 osteogenic differentiation Effects 0.000 title claims abstract description 53
- 230000001105 regulatory effect Effects 0.000 title abstract description 24
- 210000000130 stem cell Anatomy 0.000 claims abstract description 71
- 239000003814 drug Substances 0.000 claims abstract description 11
- 239000002299 complementary DNA Substances 0.000 claims abstract description 5
- 229940079593 drug Drugs 0.000 claims abstract description 5
- 125000004122 cyclic group Chemical group 0.000 claims abstract description 4
- 230000014509 gene expression Effects 0.000 claims description 35
- 108020004459 Small interfering RNA Proteins 0.000 claims description 20
- 230000000694 effects Effects 0.000 claims description 11
- 239000003112 inhibitor Substances 0.000 claims description 11
- 230000002401 inhibitory effect Effects 0.000 claims description 11
- 108091081021 Sense strand Proteins 0.000 claims description 5
- 230000000692 anti-sense effect Effects 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 210000000988 bone and bone Anatomy 0.000 abstract description 28
- 238000000034 method Methods 0.000 abstract description 17
- 230000001276 controlling effect Effects 0.000 abstract description 9
- 238000001727 in vivo Methods 0.000 abstract description 9
- 230000008929 regeneration Effects 0.000 abstract description 8
- 238000011069 regeneration method Methods 0.000 abstract description 8
- 238000002474 experimental method Methods 0.000 abstract description 6
- 238000000338 in vitro Methods 0.000 abstract description 6
- 238000005304 joining Methods 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 23
- 238000010186 staining Methods 0.000 description 22
- 230000006698 induction Effects 0.000 description 18
- 239000013642 negative control Substances 0.000 description 18
- 230000004069 differentiation Effects 0.000 description 16
- 230000002188 osteogenic effect Effects 0.000 description 15
- 108091028075 Circular RNA Proteins 0.000 description 11
- RGCKGOZRHPZPFP-UHFFFAOYSA-N alizarin Chemical compound C1=CC=C2C(=O)C3=C(O)C(O)=CC=C3C(=O)C2=C1 RGCKGOZRHPZPFP-UHFFFAOYSA-N 0.000 description 10
- 210000001519 tissue Anatomy 0.000 description 9
- 239000002609 medium Substances 0.000 description 8
- 239000008194 pharmaceutical composition Substances 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 239000004055 small Interfering RNA Substances 0.000 description 8
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 230000001737 promoting effect Effects 0.000 description 7
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 6
- 230000033228 biological regulation Effects 0.000 description 6
- 229910052791 calcium Inorganic materials 0.000 description 6
- 239000011575 calcium Substances 0.000 description 6
- 238000010586 diagram Methods 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 210000000963 osteoblast Anatomy 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 230000002018 overexpression Effects 0.000 description 5
- 238000001890 transfection Methods 0.000 description 5
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 4
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 4
- 108700024394 Exon Proteins 0.000 description 4
- 208000008589 Obesity Diseases 0.000 description 4
- 238000009825 accumulation Methods 0.000 description 4
- 210000001789 adipocyte Anatomy 0.000 description 4
- 230000011759 adipose tissue development Effects 0.000 description 4
- 230000003828 downregulation Effects 0.000 description 4
- 239000003937 drug carrier Substances 0.000 description 4
- 230000030279 gene silencing Effects 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000011580 nude mouse model Methods 0.000 description 4
- 235000020824 obesity Nutrition 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000002441 reversible effect Effects 0.000 description 4
- 230000003827 upregulation Effects 0.000 description 4
- 241000699660 Mus musculus Species 0.000 description 3
- 210000000577 adipose tissue Anatomy 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 238000001638 lipofection Methods 0.000 description 3
- 108091070501 miRNA Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 208000015580 Increased body weight Diseases 0.000 description 2
- 108091046869 Telomeric non-coding RNA Proteins 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000009815 adipogenic differentiation Effects 0.000 description 2
- 239000005557 antagonist Substances 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 210000000845 cartilage Anatomy 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 210000001612 chondrocyte Anatomy 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 230000011164 ossification Effects 0.000 description 2
- 230000001582 osteoblastic effect Effects 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 238000007363 ring formation reaction Methods 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 210000005167 vascular cell Anatomy 0.000 description 2
- 230000004580 weight loss Effects 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 102100032912 CD44 antigen Human genes 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 102100037241 Endoglin Human genes 0.000 description 1
- 108060002716 Exonuclease Proteins 0.000 description 1
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 1
- 101000881679 Homo sapiens Endoglin Proteins 0.000 description 1
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 1
- 238000012404 In vitro experiment Methods 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- NPGIHFRTRXVWOY-UHFFFAOYSA-N Oil red O Chemical compound Cc1ccc(C)c(c1)N=Nc1cc(C)c(cc1C)N=Nc1c(O)ccc2ccccc12 NPGIHFRTRXVWOY-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 238000002944 PCR assay Methods 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 208000035472 Zoonoses Diseases 0.000 description 1
- 210000000579 abdominal fat Anatomy 0.000 description 1
- 230000002293 adipogenic effect Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000009816 chondrogenic differentiation Effects 0.000 description 1
- 230000002648 chondrogenic effect Effects 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 102000013165 exonuclease Human genes 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000007443 liposuction Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007762 localization of cell Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000000107 myocyte Anatomy 0.000 description 1
- 230000003988 neural development Effects 0.000 description 1
- 108091027963 non-coding RNA Proteins 0.000 description 1
- 102000042567 non-coding RNA Human genes 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 229950003937 tolonium Drugs 0.000 description 1
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 206010048282 zoonosis Diseases 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0654—Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/01—Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
- C12N2506/13—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from connective tissue cells, from mesenchymal cells
- C12N2506/1346—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from connective tissue cells, from mesenchymal cells from mesenchymal stem cells
- C12N2506/1384—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from connective tissue cells, from mesenchymal cells from mesenchymal stem cells from adipose-derived stem cells [ADSC], from adipose stromal stem cells
Abstract
The application relates to a method for regulating and controlling osteogenic differentiation of ADSCs and tissue regeneration of Circ RNA-FTO and application thereof, belonging to the technical field of bone tissue regeneration, wherein the Circ RNA-FTO circbase ID is has-Circ-0005941, and the cDNA sequence is shown in SEQ ID NO:1 is a cyclic structure formed by joining the sequences shown in the sequence end to end. In vitro and in vivo experiments for regulating and controlling ADSCs osteogenic differentiation and tissue regeneration through circRNA-FTO prove that the circRNA-FTO plays an important role in regulating and controlling the osteogenic differentiation and/or tissue regeneration of adipose-derived stem cells; the Circ RNA-FTO can be used as a potential target point of tissue regeneration treatment, so that new relevant medicines or factors related to the Circ RNA-FTO are promoted to be developed, and finally the medicines or factors are applied to the field of regeneration medicine, thereby achieving real accurate medical treatment in clinic.
Description
Technical field:
the application belongs to the technical field of bone tissue regeneration, and particularly relates to a Circ RNA-FTO for regulating and controlling ADSCs osteogenic differentiation and tissue regeneration and application thereof.
The background technology is as follows:
bone defects caused by trauma, tumors and infection are common clinical diseases, and autologous bone grafting has been the treatment of choice for bone defects, but is greatly limited due to its limited sources, poor plasticity, and the potential for donor site injury. Allograft bone grafting presents immune rejection. If the treatment to remove cellular components is performed in order to reduce the immunogenicity of the allograft bone, it may promote its loss of osteogenic activity. The source is limited and the infection rate is high. The immune rejection of xenogeneic bone grafts is more severe and can infect zoonotic diseases. They create a heavy social and economic burden, severely affecting the quality of life of the patient. Adipose stem cells (adipose Derived Stem Cells, ADSCs) have attracted considerable attention in recent years. The adipose-derived stem cells have wide sources, more content, simple acquisition mode and less pain born by patients. Under appropriate circumstances, adipose stem cells can differentiate into adipocytes, osteoblasts, chondroblasts, myocytes, and the like. ADSCs have a multidirectional differentiation capacity and a relatively clear osteogenic potential, and therefore become important seed cells in bone tissue engineering.
The circular RNA (circRNA) is one of non-coding RNAs, and is another new research hotspot of RNA families following micro RNAs (mirnas) and long non-coding RNAs (lncrnas). The circRNA consists of a covalent closed-loop structure without a 5 'cap or 3' polyadenylation tail. Compared to linear RNAs, circRNA has the following features: high stability; high degree of evolutionary conservation; resistant to digestion by exonuclease and not easy to degrade; most of the circRNA is localized to the cytoplasm, and a few of the circRNA plays a role in the nucleus; a tissue cell or a certain period of a cell may specifically express a specific circrna. The ability of circular RNAs to exhibit developmental stage or tissue-specific expression makes them ideal biomarkers for disease diagnosis and targeted therapy. Several studies explored the important role of circRNA in the development of cellular activity, embryonic development, neural development, and various human diseases.
However, studies on the action of Circ RNA-FTO in adipose-derived stem cells and the actions related to the osteogenic differentiation and tissue regeneration of adipose-derived stem cells have not been reported.
The application comprises the following steps:
aiming at the prior art, the inventor firstly proposes a new concept of utilizing adipose-derived stem cells to regenerate bone tissues, and carries out series of related in-vivo and in-vitro experiments, such as successful regeneration of bone tissues on a nude mouse ectopic osteogenesis and rabbit mandibular defect animal model; the autologous fat stem cells are attached with the biological scaffold to be used for tissue regeneration, so that a good effect is obtained. Further, by researching in vitro and in vivo experiments of regulating ADSCs osteogenic differentiation and tissue regeneration by using the circRNA-FTO, molecular level research and in vivo tissue regeneration research are carried out, and the result shows that the circRNA-FTO plays an important role in regulating and controlling the adipose-derived stem cell osteogenic differentiation and tissue regeneration, and by reverse demonstration, when the circRNA-FTO is downregulated, the osteogenic differentiation and bone tissue regeneration capacity of the ADSCs is inhibited, and the application clarifies the regulating and controlling function of the circRNA-FTO in adipose-derived stem cell mediated tissue regeneration, thereby providing an important basis for regulating and controlling adipose-derived stem cells to promote bone tissue regeneration.
In order to achieve the above purpose, the present application adopts the following technical scheme:
regulating osteogenic differentiation of ADSCs and tissue regeneration of circRNA, specifically circRNA-FTO, with circbase ID of hsa-Circ-0005941.
The cDNA sequence of the Circ RNA-FTO is shown as SEQ ID NO:1 is a cyclic structure formed by joining the sequences shown in the sequence end to end.
The application of the circRNA-FTO in the regulation and control of the osteogenic differentiation and the tissue regeneration of ADSCs comprises positive regulation and negative regulation, wherein the positive regulation refers to the promotion of the osteogenic differentiation and the tissue regeneration of the adipose-derived stem cells, and the negative regulation refers to the inhibition of the osteogenic differentiation and the tissue regeneration of the adipose-derived stem cells.
The application of the circRNA-FTO in the regulation and control of the osteogenic differentiation and the tissue regeneration of ADSCs improves the expression of the circRNA-FTO and promotes the osteogenic differentiation and the tissue regeneration of adipose-derived stem cells; the expression of Circ RNA-FTO is reduced, and the osteogenic differentiation of ADSCs and the regeneration capacity of bone tissue are inhibited.
Use of a Circ RNA-FTO promoter in the preparation of a medicament for promoting osteoblastic differentiation and tissue regeneration of adipose-derived stem cells.
The circle RNA-FTO promoter includes any substance that can increase the activity of circle RNA-FTO and increase the expression level of circle RNA-FTO, such as an over-expression vector of circle RNA-FTO that increases the expression level of circle RNA-FTO.
Use of a Circ RNA-FTO inhibitor for the preparation of a medicament for inhibiting adipose stem cell osteogenic differentiation and tissue regeneration.
The Circ RNA-FTO inhibitor includes any substance that can reduce the activity of Circ RNA-FTO and reduce the expression level of Circ RNA-FTO, and specifically includes antagonists, downregulators, blockers, etc.
The circRNA-FTO inhibitor is a small interfering RNA sequence for inhibiting the expression of the circRNA-FTO.
The sir target sequence of the Circ RNA-FTO is as follows:
sense strand: GGAGGGUGUGAUGAUCUCAUU
Antisense strand: UGAGAUCAUCACACCCUCCAA.
A method of promoting adipose stem cell osteogenic differentiation and bone tissue regeneration, the method comprising increasing the activity of and/or increasing the expression of Circ RNA-FTO.
The method for promoting the osteogenic differentiation of the adipose-derived stem cells and the regeneration of the bone tissues further comprises the step of preparing the adipose-derived stem cells into an osteogenic material by forming an adipose-derived stem cell complex with an artificial bone scaffold material by improving the activity of the circRNA-FTO and/or increasing the expression of the circRNA-FTO.
A method of inhibiting adipose stem cell osteogenic differentiation and bone tissue regeneration, the method comprising inhibiting expression of Circ RNA-FTO.
The Circ RNA-FTO is used as a target spot for non-disease diagnosis and treatment and is applied to regulation and control of the osteogenic differentiation and tissue regeneration of adipose-derived stem cells.
The adipose-derived stem cells have various differentiation potential, can form various tissues such as bones, cartilages, nerves, fat, blood vessels and the like through proper induction in vitro, can be accompanied with the generation of various differentiation cells (osteoblast + adipocyte, osteoblast + vascular cell and the like) during induction (even in the directional induction process), can effectively induce the differentiation of stem cells to other specific tissue directions through partial inhibition of bone differentiation.
A pharmaceutical composition for modulating osteogenic differentiation and tissue regeneration of adipose stem cells, said pharmaceutical composition comprising an effective amount of the above-described circrna-FTO promoter or circrna-FTO inhibitor.
The pharmaceutical composition for regulating the osteogenic differentiation and the tissue regeneration of the adipose-derived stem cells further comprises: a pharmaceutically acceptable carrier.
The pharmaceutically acceptable carrier in the pharmaceutical composition for regulating the osteogenic differentiation and the tissue regeneration of the adipose-derived stem cells is a conventional administration carrier, and comprises various excipients or diluents and the like.
The Circ RNA-FTO of the application is derived from chr16:53907697-53922863, has a gene length of 15166 bases, is formed by cyclization of 5,6,7 exons of the FTO, and has a ring length of 334 nucleotides. FTO is a very large gene whose 9 exons span over 400kb on chromosome 16. Fat mass and obesity-related FTO is reported to be one of the key factors in regulating body weight and fat mass, and it is involved in regulating common obesity and body mass index. FTO participates in fat accumulation primarily by regulating adipogenesis during adipogenesis. Inhibition of FTO or FTO mutations has been reported to result in weight loss and fat accumulation reduction. In contrast, overexpression of FTO results in increased body weight and fat mass.
The application has the beneficial effects that:
in-vitro and in-vivo experiments for regulating and controlling ADSCs osteogenic differentiation and tissue regeneration by using the circRNA-FTO prove that the circRNA-FTO plays an important role in regulating and controlling the osteogenic differentiation and/or tissue regeneration of adipose-derived stem cells; the Circ RNA-FTO can be used as a potential target point of tissue regeneration treatment, so that new relevant medicines or factors related to the Circ RNA-FTO are promoted to be developed, and finally the medicines or factors are applied to the field of regeneration medicine, thereby achieving real accurate medical treatment in clinic.
Description of the drawings:
FIG. 1 is a flow chart of the detection of hADSCs cell surface markers of example 1;
FIG. 2 is a diagram showing the multidirectional differentiation of hADSCs of example 1, from left to right: hADSCs cultivated to passage 3; adipogenic differentiation of cultured hADSCs; osteogenic differentiation of hADSCs; chondrogenic differentiation of hADSCs;
FIG. 3 is a graph showing the results of alizarin red staining of adipose-derived stem cells after osteogenic induction of example 1; the steps from left to right are as follows: alizing the adipose-derived stem cells for 7 days, 14 days and 21 days, and performing alizarin red staining;
FIG. 4 is a graph showing the RT-q PCR assay of example 2 for the variation of expression of Circ RNA-FTO 0, 3, 5 and 7 days after osteogenic induction of adipose-derived stem cells;
FIG. 5 is a fluorescence microscopy image of the silencing efficacy of the small interfering RNA transfected nc control and the Circ RNA-FTO experimental group of example 2;
FIG. 6 is a graph showing the expression of the Circ RNA-FTO in the nc control group and the experimental group by RT-PCR in example 2;
FIG. 7 is a Western blot detection of relative expression of osteogenic differentiation-related proteins after osteogenic induction of hADSCs transfected by siRNA-NC group and siRNA-FTO group, respectively, in example 2;
FIG. 8 is a graph showing the number and extent of osteogenic calcium nodules detected by alkaline phosphatase staining and alizarin red staining of hADSCs transfected with siRNA-NC and siRNA-FTO groups, respectively, of example 2, wherein alkaline phosphatase staining is the upper one and alizarin red staining is the lower one;
FIG. 9 is a graph showing the change in the in vivo tissue regeneration ability after HE staining and Masson staining, respectively, of hADSCs+Bio-Os transfected with siRNA circFTO and siRNA NC of example 2.
The specific embodiment is as follows:
it is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present application. As used herein, the singular forms also are intended to include the plural forms unless the context clearly indicates otherwise, and furthermore, it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, and/or combinations thereof.
In order to enable those skilled in the art to more clearly understand the technical scheme of the present application, the technical scheme of the present application will be described in detail with reference to specific embodiments.
Regulating osteogenic differentiation of ADSCs and tissue regeneration of circRNA, specifically circRNA-FTO, with circbase ID of hsa-Circ-0005941.
The cDNA sequence of the Circ RNA-FTO is shown as SEQ ID NO:1 is a cyclic structure formed by joining the sequences shown in the sequence end to end.
The application of the circRNA-FTO in the regulation and control of the osteogenic differentiation and the tissue regeneration of ADSCs comprises positive regulation and negative regulation, wherein the positive regulation refers to the promotion of the osteogenic differentiation and the tissue regeneration of the adipose-derived stem cells, and the negative regulation refers to the inhibition of the osteogenic differentiation and the tissue regeneration of the adipose-derived stem cells.
The application of the circRNA-FTO in the regulation and control of the osteogenic differentiation and the tissue regeneration of ADSCs improves the expression of the circRNA-FTO and promotes the osteogenic differentiation and the tissue regeneration of adipose-derived stem cells; the expression of Circ RNA-FTO is reduced, and the osteogenic differentiation of ADSCs and the regeneration capacity of bone tissue are inhibited.
Use of a Circ RNA-FTO promoter in the preparation of a medicament for promoting osteoblastic differentiation and tissue regeneration of adipose-derived stem cells.
The circle RNA-FTO promoter includes any substance that can increase the activity of circle RNA-FTO and increase the expression level of circle RNA-FTO, such as an over-expression vector of circle RNA-FTO that increases the expression level of circle RNA-FTO.
Use of a Circ RNA-FTO inhibitor for the preparation of a medicament for inhibiting adipose stem cell osteogenic differentiation and tissue regeneration.
The Circ RNA-FTO inhibitor includes any substance that can reduce the activity of Circ RNA-FTO and reduce the expression level of Circ RNA-FTO, and specifically includes antagonists, downregulators, blockers, etc.
The circRNA-FTO inhibitor is a small interfering RNA sequence for inhibiting the expression of the circRNA-FTO.
The sir target sequence of the Circ RNA-FTO is as follows:
sense strand: 5'-ggagggugugaugaucucauu-3'
Antisense strand: 5'-ugagaucaucacacccuccaa-3'
A method of promoting adipose stem cell osteogenic differentiation and bone tissue regeneration, the method comprising increasing the activity of and/or increasing the expression of Circ RNA-FTO.
The method for promoting the osteogenic differentiation of the adipose-derived stem cells and the regeneration of the bone tissues further comprises the step of preparing the adipose-derived stem cells into an osteogenic material by forming an adipose-derived stem cell complex with an artificial bone scaffold material by improving the activity of the circRNA-FTO and/or increasing the expression of the circRNA-FTO.
A method of inhibiting adipose stem cell osteogenic differentiation and bone tissue regeneration, the method comprising inhibiting expression of Circ RNA-FTO.
The Circ RNA-FTO is used as a target spot for non-disease diagnosis and treatment and is applied to regulation and control of the osteogenic differentiation and tissue regeneration of adipose-derived stem cells.
The adipose-derived stem cells have various differentiation potential, can form various tissues such as bones, cartilages, nerves, fat, blood vessels and the like through proper induction in vitro, can be accompanied with the generation of various differentiation cells (osteoblast + adipocyte, osteoblast + vascular cell and the like) during induction (even in the directional induction process), can effectively induce the differentiation of stem cells to other specific tissue directions through partial inhibition of bone differentiation.
A pharmaceutical composition for modulating osteogenic differentiation and tissue regeneration of adipose stem cells, said pharmaceutical composition comprising an effective amount of the above-described circrna-FTO promoter or circrna-FTO inhibitor.
The pharmaceutical composition for regulating the osteogenic differentiation and the tissue regeneration of the adipose-derived stem cells further comprises: a pharmaceutically acceptable carrier.
The pharmaceutically acceptable carrier in the pharmaceutical composition for regulating the osteogenic differentiation and the tissue regeneration of the adipose-derived stem cells is a conventional administration carrier, and comprises various excipients or diluents and the like.
The Circ RNA-FTO of the application is derived from chr16:53907697-53922863, has a gene length of 15166 bases, is formed by cyclization of 5,6,7 exons of the FTO, and has a ring length of 334 nucleotides. FTO is a very large gene whose 9 exons span over 400kb on chromosome 16. Fat mass and obesity-related FTO is reported to be one of the key factors in regulating body weight and fat mass, and it is involved in regulating common obesity and body mass index. FTO participates in fat accumulation primarily by regulating adipogenesis during adipogenesis. Inhibition of FTO or FTO mutations has been reported to result in weight loss and fat accumulation reduction. In contrast, overexpression of FTO results in increased body weight and fat mass.
EXAMPLE 1 biological Properties of adipose Stem cells
1. Isolation and culture of human adipose-derived stem cells
Abdominal adipose tissue was taken during liposuction and stored in 20mL sterile syringes for later use and ice bags. The adipose tissue was centrifuged at 1000rpm/5min in a 50mL centrifuge tube and the underlying liquid was removed. Adipose tissue was treated with 1:1 to 0.2% type I collagenase was digested by shaking table at 37℃for 40 minutes and was seen to separate into three layers after standing. The upper grease layer was removed and thoroughly shaken and filtered (200 mesh screen). The supernatant was then centrifuged three times at 1000rmp/5min to remove the supernatant, and the cell pellet was visible. The cells were blown uniformly with the cell culture solution. Cells were then seeded into petri dishes at a density of 5X 105/mL. The cells were then placed in 5% CO at 37℃ 2 In an incubator at 95% humidity. And (5) subculturing to the third generation to obtain the third generation adipose-derived stem cells hADSCs. (third generation cells complete subsequent experiments)
2. Expression of markers on the surface of adipose-derived stem cells
The expression of mesenchymal stem cell markers such as FITC-CD73, CD44, CD105, CD45 and the like of the adipose stem cells is observed by using flow cytometry and immunofluorescence. According to the national institute of medical biotechnology (TCM) on the mass management expert consensus on the extraction, preparation and storage of adipose tissue-derived stem cells, the experimental cells conform to the surface marking characteristics of mesenchymal stem cells.
3. Multidirectional differentiation ability
Culturing the third-generation human adipose-derived stem cells in the step 1 in an osteogenic induction medium, an adipogenic induction medium and a chondrogenic induction medium for 3 weeks respectively, and detecting the multidirectional differentiation potential of the human adipose-derived stem cells through alizarin red staining, oil red O staining and toluidine blue staining. The flow detection diagram of the hADSCs cell surface markers is shown in FIG. 1, the multi-directional differentiation diagram of the hADSCs is shown in FIG. 2, and the flow detection diagram is from left to right: hADSCs cultivated to passage 3; adipogenic differentiation of cultured hADSCs; osteogenic differentiation of hADSCs; the isolated and cultured hADSCs can be differentiated into osteoblasts, adipocytes and chondroblasts through identification and induction, and have the multi-directional differentiation potential of stem cells. The result of alizarin red staining of adipose-derived stem cells after osteogenic induction is shown in FIG. 3; the steps from left to right are as follows: the results of alizing the adipose-derived stem cells for 7 days, 14 days and 21 days were stained with alizing red alizing medium to induce hADSCs for 7 days, 14 days and 21 days. The number of calcium nodules in 7 days is small, the volume is small, the coloring is light, and the distribution is scattered. The number of calcium nodules in the group is obviously increased in 14 days compared with 7 days, the volume is increased, and a plurality of calcium nodules can be gathered into clusters. Alizarin red staining of the hADSCs at 21 days was significantly stronger than that induced by the hADSCs at 14 days, with the visual field filled with calcium nodules.
EXAMPLE 2 selection of Circ RNA-FTO and cell localization and expression Change in osteogenic Induction thereof
1. And respectively performing osteogenic induction on the adipose-derived stem cells for 0d, 3d, 5d and 7d, extracting RNA, and detecting the expression change trend of the circRNA-FTO at different times by RT-q PCR.
The experimental results are shown in FIG. 4, in which the expression of circRNA-FTO increases with time during the osteogenic differentiation.
2. Design and transfection of small interfering RNA sequences
The Circ RNA-FTO sequence (circbase ID: hsa-Circ-0005941) was designed as a primer synthesized and purified by Shanghai Ji Ma Biotechnology Co., ltd, and a small interfering RNA sequence was designed to inhibit the expression of Circ RNA-FTO (because of the excessive size of the FTO sequence, it is extremely difficult to successfully construct an over-expression vector, and thus the application was demonstrated by reverse research by inhibiting the expression of Circ RNA-FTO).
According to the starting point and the ending point of the human circRNA-FTO sequence as the splicing point, designing the thermal stability based on the siRNA near the splicing point, selecting the base at the tail, designing the siRNA optimal target point of the circRNA by GC content, and selecting the optimal 1 corresponding target point sequences as follows:
sense strand: 5'-ggagggugugaugaucucauu-3'
Antisense strand: 5'-ugagaucaucacacccuccaa-3'
The negative control NC (Negative Control) sequence is as follows:
sense strand: 5'-uucuccgaacgugucacguuu-3'
Antisense strand: 5'-acgugacacguucggagaauu-3'
Grouping cells: siRNA-NC group and siRNA-FTO group, respectively transferred into hADSCs by liposome transfection method and Negative Control (NC). The experiment sets up an experimental group and a negative control group. The experimental group is cells transfected with siRNA; the negative control group was NC control cells. After 12h, the medium is changed back to normal medium, and after 24h incubation, the medium is observed under a fluorescence microscope, and green fluorescence appears to indicate that transfection is successful. Extracting total RNA and completing the related verification of the subsequent RT-q PCR.
The detection results of transfection and silencing efficiency show that siRNA transfected hADSC cells can obviously inhibit the expression of Circ RNA-FTO, a fluorescent microscope diagram for detecting silencing efficiency of a small interfering RNA transfected nc control group and a Circ RNA-FTO experimental group is shown in FIG. 5, and a diagram for detecting the expression of the Circ RNA-FTO of the nc control group and the experimental group by RT-PCR is shown in FIG. 6;
3. detection of osteogenic related proteins and osteogenic related staining by silencing Circ RNA-FTO
Cells were washed with serum-free medium and siRNA-NC and siRNA-FTO groups were added to the cells according to the transfection procedure instructions. After 12h, the culture medium is changed into normal culture medium, and the culture medium is incubated for 72h, and after transfection is stabilized, the culture medium is used for osteogenesis induction. The subsequent functional study of the Circ RNA-FTO on the osteogenic differentiation of the adipose-derived stem cells is completed.
The detection experimental results of relative expression of osteogenic differentiation related proteins of ADSCs transfected by the siRNA-NC group and the siRNA-FTO group respectively are shown in a graph in FIG. 7, and the expression of the osteogenic differentiation related factors (ALP, runx2 and OCN) of the siRNA-FTO group is reduced compared with that of the siRNA-NC group, so that the circRNA-FTO has the osteogenic differentiation capability, and the quantity and degree graphs of osteogenic differentiation calcium nodules detected by the alkaline phosphatase staining and alizarin red staining of the ADSCs transfected by the siRNA-NC group and the siRNA-FTO group respectively are shown in a graph in FIG. 8, wherein the alkaline phosphatase staining is the upper part, and the alizarin red staining is the lower part; the results show that ALP staining and alizarin red staining show that the siRNA-NC group and the siRNA-FTO group have obvious differences after the osteogenic induction of the hADSCs.
EXAMPLE 3 study of Circ RNA-FTO to regulate tissue regeneration in adipose-derived stem cells
Human adipose stem cell complexes were implanted subcutaneously in the back of nude mice and their tissue forming ability was observed.
1. siRNA and Negative Control (NC) were transferred into hADSCs by liposome transfection
Human adipose-derived stem cells were cultured by a tissue mass method and an enzyme digestion method, and third generation adipose-derived stem cells which grew vigorously were inoculated into a 60mm dish with an alpha-MEM medium (containing 10% fetal bovine serum, 2mmol/L glutamine, 100U/ml penicillin, 100. Mu.g/ml streptomycin). siRNA and Negative Control (NC) were transferred into adipose-derived stem cells by liposome transfection, and the experiments were divided into 2 groups: (1) siRNA-NC transfected adipose-stem cell group (2) siRNA-FTO transfected adipose-stem cell group.
2. Preparation of biological scaffold material
The autoclaved Bio-Oss collagen scaffold material was co-cultured with adipose stem cells cultured in vitro in a ratio of 50mg:1*10 7 Individual cells. The randomization was divided into 2 groups: (1) siRNA-NC transfected adipose-derived stem cells+Bio-Oss group (2) siRNA-FTO transfected adipose-derived stem cells+Bio-Oss group.
3. In vivo implantation of scaffold material/stem cell complexes
Nude mice were randomly divided into 2 groups: (1) siRNA NC transfected hADSCs+Bio-Os group; (2) siRNA circFTO transfected hADSCs+Bio-Os group; after 8w of the back planting, the nude mice are sacrificed to obtain specimens, fixed, decalcified, HE and Masson staining are carried out, and the regeneration condition of the biological stent tissue is observed.
As shown in FIG. 9, the variation of the in vivo tissue regeneration capacity after the circRNA-FTO is obviously reduced (H & E staining and Masson staining show after 8w in vivo), the bone tissue content of the siRNA-FTO transfected adipose-stem cells+Bio-Os group is very small, and the circRNA-FTO has a promoting effect on the tissue regeneration capacity.
The above examples are preferred embodiments of the present application, but the embodiments of the present application are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present application should be made in the equivalent manner, and the embodiments are included in the protection scope of the present application.
Sequence listing
<110> Liaoning province tumor Hospital
<120> modulation of ADSCs osteogenic differentiation and tissue regeneration circRNA-FTO and uses thereof
<160> 5
<170> SIPOSequenceListing 1.0
<210> 30
<211> 1518
<212> DNA
<213> cDNA sequence of hsa-circ-0005941 Gene (Unknown)
<400> 30
atgaagcgaa ccccaaccgc cgaggaacga gagcgcgaag ctaagaaact gaggcttctt 60
gaagagctgg aagacacttg gcttccttat ctgaccccca aagatgatga attctatcag 120
cagtggcagc tgaaataccc taagctaatt ctccgagaag caggcagcgt ccctgaggga 180
ctccacaaag aggttcaaga agccttcctc gcactgcaca agcatggctg cttatttcgg 240
gacctggtca ggatccaagg caaagatttg ctcacgccag tatctcgcct cctcattggt 300
aaccccggct gcacctacaa gtacctgaac accaggctct tcacggtccc ctggccagtg 360
aagggctctg atgcaaagta caatgaggcc gagataggcg ccgcctgcca gaccttcctc 420
aagctcaacg actacctgca gattgagacc atccaggcgc tggaggaact cgctgccaag 480
gagaaagcca atatcgacac cgtgccggtg tgtataggtc cagatttccc cagggtcggc 540
atggggtcat cctttgacgg gcatgacgag gtggacagga agagcagagc cgcctacaac 600
ctaactttgt tgaacttcat ggatccccag aaaatgccgt acctgaaaga ggagccctac 660
tttggcatgg ggaagatggc tgtgagctgg catcacgatg aaaatctggt ggacaggtca 720
gcggtggcag tgtacaatta tagctgtgaa ggccctgaag aggaaagcga ggatgatccc 780
cagctcgaag gcagagatcc cgatgtgtgg catgttggct ttaagatctc atgggacata 840
gagacccctg gtttggcgat accccttcac caaggagact gctactttat gctggatgat 900
ctcaatgcca cccaccaaca ctgtgttttg gctggtttac caccccggtt tagttccacc 960
caccgagtgg ccgagtgctc gacgggaacc ttggattaca tcttacagcg ctgccagttg 1020
gccctgcaga atgtccgtga tgaggcggac agtggtgaag tctctttgaa atccttggag 1080
cctgcggttt tgaaacaagg agaagaaatc cacaacgagg tcgagtttga gtggctgaga 1140
cagttttggt ttcaaggcaa tcgatacaaa aagtgcaccg attggtggtg tcaacccatg 1200
actcagctgg aagagctttg gaagaagatg gaaggtgcga cccatgctgt gcttcgtgaa 1260
gttaggagag agggggcccc tgtggaacag agcagtgaca tcctgactgc catcctagcc 1320
gtgctcacca ctcgccagaa cctgaggagg gagtggcatg ccaggtgcca gtcccgaatt 1380
gcccgaactc tgcctgtgga ccagaagcca gaatgccggc cgtattggga aaaggatgat 1440
ccctccatgc ctctgccgtt tgatctcaca gacactgtgg ctgaactcag aggtctgctt 1500
ctggaagcca aaccctag 1518
<210> 29
<211> 21
<212> RNA
<213> hsa-circ-0005941 Small interfering RNA sequence Forward primer (Unknown)
<400> 29
ggagggugug augaucucau u 21
<210> 30
<211> 21
<212> RNA
<213> hsa-circ-0005941 Small interfering RNA sequence reverse primer (Unknown)
<400> 30
ugagaucauc acacccucca a 21
<210> 29
<211> 21
<212> RNA
<213> negative control NC sequence Forward primer (Unknown)
<400> 29
uucuccgaac gugucacguu u 21
<210> 30
<211> 21
<212> RNA
<213> negative control NC sequence reverse primer (Unknown)
<400> 30
acgugacacg uucggagaau u 21
Claims (1)
1. The application of the inhibitor of the Circ RNA-FTO is characterized in that the cDNA sequence of the Circ RNA-FTO is shown as SEQ ID NO:1 is a cyclic structure formed by connecting sequences shown in the sequence end to end; the circRNA-FTO inhibitor is used for preparing medicines for inhibiting the osteogenic differentiation and tissue regeneration of adipose stem cells, and comprises substances capable of reducing the activity of the circRNA-FTO and reducing the expression quantity of the circRNA-FTO;
the Circ RNA-FTO inhibitor is an siRNA sequence for inhibiting the expression of the Circ RNA-FTO, and the siRNA target sequence of the Circ RNA-FTO is as follows:
sense strand: 5'-ggagggugugaugaucucauu-3'
Antisense strand: 5'-ugagaucaucacacccuccaa-3'.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010321971.1A CN111500578B (en) | 2020-04-22 | 2020-04-22 | Regulating ADSCs osteogenic differentiation and tissue regeneration circRNA-FTO and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010321971.1A CN111500578B (en) | 2020-04-22 | 2020-04-22 | Regulating ADSCs osteogenic differentiation and tissue regeneration circRNA-FTO and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111500578A CN111500578A (en) | 2020-08-07 |
| CN111500578B true CN111500578B (en) | 2023-12-12 |
Family
ID=71865957
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010321971.1A Active CN111500578B (en) | 2020-04-22 | 2020-04-22 | Regulating ADSCs osteogenic differentiation and tissue regeneration circRNA-FTO and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111500578B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112458095A (en) * | 2020-11-30 | 2021-03-09 | 辽宁省肿瘤医院 | LMO3 gene for regulating and controlling osteogenic differentiation and tissue regeneration of ADSCs and application thereof |
| CN113151279B (en) * | 2021-05-12 | 2023-05-23 | 中山大学附属第一医院 | hsa_circ_0001236 and application thereof |
| CN113308438B (en) * | 2021-06-11 | 2022-09-16 | 浙江大学 | FTO gene modified porcine adipose-derived stem cell and construction method and application thereof |
| CN115141808A (en) * | 2022-06-21 | 2022-10-04 | 中山大学附属第八医院（深圳福田） | Hsa _ circ _0001493 and hsa _ circ _0070562 inhibit osteogenic differentiation |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3237642A4 (en) * | 2014-12-24 | 2018-09-19 | Massachusetts Institute of Technology | Compositions and methods for manipulation of adipocyte energy consumption regulatory pathway |
-
2020
- 2020-04-22 CN CN202010321971.1A patent/CN111500578B/en active Active
Non-Patent Citations (2)
| Title |
|---|
| Chen,X.等.GenBank:KM232950.1.《GenBank》.2018,第1-2页. * |
| Yue Kang等.Differential circular RNA expression profiling during osteogenic differentiation in human adipose-derived stem cells.《Epigenomics》.2020,第12卷(第4期),摘要、附加材料表2. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111500578A (en) | 2020-08-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111500578B (en) | Regulating ADSCs osteogenic differentiation and tissue regeneration circRNA-FTO and application thereof | |
| Hu et al. | Stem cell‐based tooth and periodontal regeneration | |
| Deng et al. | The role of miR-31-modified adipose tissue-derived stem cells in repairing rat critical-sized calvarial defects | |
| Charbord | Bone marrow mesenchymal stem cells: historical overview and concepts | |
| Gong et al. | Current advance and future prospects of tissue engineering approach to dentin/pulp regenerative therapy | |
| Liao et al. | Osteogenic differentiation of adipose-derived stem cells and calvarial defect repair using baculovirus-mediated co-expression of BMP-2 and miR-148b | |
| Wang et al. | Promotion of cardiac differentiation of brown adipose derived stem cells by chitosan hydrogel for repair after myocardial infarction | |
| Tian et al. | Myogenic differentiation of human bone marrow mesenchymal stem cells on a 3D nano fibrous scaffold for bladder tissue engineering | |
| Hu et al. | Neuronally differentiated adipose-derived stem cells and aligned PHBV nanofiber nerve scaffolds promote sciatic nerve regeneration | |
| CN1227356C (en) | Application of substrate cell derived from fat tissue | |
| KR101389850B1 (en) | Method for Culture of cardiac progenitor cells and Uses Therefor | |
| Gu | Progress and perspectives of neural tissue engineering | |
| CN107354127B (en) | Effect of the LncRNA-TUG1 in regulation PDLSCs Osteoblast Differentiation and regeneration | |
| Xu et al. | Intercellular adhesion molecule-1 inhibits osteogenic differentiation of mesenchymal stem cells and impairs bio-scaffold-mediated bone regeneration in vivo | |
| Liu et al. | The role of MicroRNAs in tendon injury, repair, and related tissue engineering | |
| Yan et al. | BMP7‑overexpressing bone marrow‑derived mesenchymal stem cells (BMSCs) are more effective than wild‑type BMSCs in healing fractures | |
| Nasser et al. | Current situation and future of stem cells in cardiovascular medicine | |
| CN1778905A (en) | Separating culture and use for fatty mesenchymal dry cell | |
| KR101503939B1 (en) | Method for preparation of cartilage cell | |
| Yin et al. | Cell‐Based Therapies for Degenerative Musculoskeletal Diseases | |
| Wu et al. | Repair of cartilage defects in BMSCs via CDMP1 gene transfection | |
| CN105878264B (en) | MiR-21 and its inhibitor are preparing the application in promoting paradenlal tissue regeneration drug | |
| Huang et al. | Calcitonin gene-related peptide-induced calcium alginate gel combined with adipose-derived stem cells differentiating to osteoblasts | |
| KR101389851B1 (en) | Method for Culture of Neural Crest Stem Cells and Uses Therefor | |
| CN113430171B (en) | Cell patch for transfecting miRNA and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |